How is Trehalulose Formed by Australian Stingless Bees? - An Intermolecular Displacement of Nectar Sucrose.

Trehalulose, a rare sucrose isomer, is a dominant sugar in stingless bee honey, with traces of the trisaccharide erlose. Incubating sucrose solutions with macerated stingless bee parts (head, thorax, and abdomen) from Tetragonula carbonaria, we observed that sucrose isomerization occurs predominantly in the head incubations, with trehalulose constituting 76.2-80.0% of total detected sugar. By contrast, sucrose hydrolysis occurred in stingless bee abdomen incubations, with glucose and fructose observed as 48.6-51.7% and 48.3-49.7%, respectively, of total detected sugar. Incubating glucose/fructose (1:1) solutions with any bee part did not result in trehalulose formation. In addition, by tracing the 13C isotope-labeled monosaccharide moieties throughout the isomerization from sucrose to trehalulose and erlose, for the first time, the mechanism was established as an enzymatic double displacement reaction. Sucrose acts as a glucose donor giving a ß-d-glucosyl enzyme intermediate with fructose release as demonstrated by mixed isotope products. Glucosylation of fructose (inter- or intramolecularly) with isomerization forms trehalulose (favorable), while glucosylation of sucrose forms erlose (less favorable).

Unique physicochemical properties and rare reducing sugar trehalulose mandate new international regulation for stingless bee honey.

Stingless bee honey (SBH) of four stingless bee species (Heterotrigona itama, Geniotrigona thoracica, Tetragonula carbonaria, and Tetragonula hockingsi) from two geographic regions (Malaysia and Australia, n = 36) were studied for their physicochemical parameters, including total phenolic and multi-elemental contents. Sugar analysis confirmed the prominent presence of trehalulose in all samples. All SBH failed to meet the CODEX Standard for honey moisture, free acidity, and total fructose plus glucose levels. One-way ANOVA, principal component analysis (PCA) and hierarchical component analysis (HCA) confirm distinctive differences between Australian and Malaysian SBH with Australian SBH having significantly (P < 0.05) higher pH, total phenolic, total mineral, and electrical conductivity attributes. Toxic elements in all SBH were much lower than standard CODEX limit. The species G. thoracica produced honey with significantly (P < 0.05) higher trehalulose than both Australian bee species. This research will add value to SBH and hasten new international standards ensuring commercial viability, safety, quality, and marketability of SBH.

Feeding Sugars to Stingless Bees: Identifying the Origin of Trehalulose-Rich Honey Composition.

The beneficial disaccharide, trehalulose, is a feature of stingless bee honey, while not dominant in any other foods. By experimentally feeding sugar solutions to confined colonies of the Australian stingless bee Tetragonula carbonaria, the origin of trehalulose has now been established. Complete conversion of fed sucrose was observed, by analysis of the honey, forming trehalulose (64-72%) with lesser erlose (18-23%), fructose (9-12%), and minor glucose detected. Remarkably, feeding solutions of glucose/fructose (1:1) mixtures did not result in trehalulose/erlose formation. Hence, stingless bees with natural access to floral nectar high in sucrose will produce honey high in trehalulose, with its associated beneficial properties. Any temptation to artificially increase trehalulose content by feeding sucrose to stingless bees would produce "fake" honey lacking key natural phytochemicals available to the foraging bee. The sucrose-fed fake and natural honey were however readily distinguished via isotope ratio mass spectrometry δ13C values, to combat such potential indirect adulteration.

Isotopic and Elemental Composition of Roasted Coffee as a Guide to Authenticity and Origin.

This study presents the stable isotopic and elemental compositions of single-origin, roasted coffees available to retail consumers. The δ(13)C, δ(15)N, and δ(18)O compositions were in agreement with those previously reported for green coffee beans. The δ(15)N composition was seen to be related to organic cultivation, reflected in both δ(2)H and δ(18)O compositions. The δ(13)C composition of extracted caffeine differed little from that of the bulk coffee. Stepwise discriminant analysis with jackknife tests, using isotopic and elemental data, provided up to 77% correct classification of regions of production. Samples from Africa and India were readily classified. The wide range in both isotopic and elemental compositions of samples from other regions, specifically Central/South America, resulted in poor discrimination between or within these regions. Simpler X-Y and geo-spatial plots of the isotopic data provided effective visual means to distinguish between coffees from different regions.

Stable isotope and chemical compositions of European and Australasian ciders as a guide to authenticity.

This paper presents a data set derived from the analysis of bottled and canned ciders that may be used for comparison with suspected counterfeit or substitute products. Isotopic analysis of the solid residues from ciders (predominantly sugar) provided a means to determine the addition of C4 plant sugars. The added sugars were found to comprise cane sugar, high-fructose corn syrup, glucose, or combinations. The majority of ciders from Australia and New Zealand were found to contain significant amounts of added sugar, which provided a limited means to distinguish these ciders from European ciders. The hydrogen and oxygen isotopic compositions of the whole ciders (predominantly water) were shown to be controlled by two factors, the water available to the parent plant and evaporation. Analysis of data derived from both isotopic and chemical analysis of ciders provided a means to discriminate between regions and countries of manufacture.